6alpha-fluoro delta-cortisone and 6alpha-fluoro delta&#39;-hydrocortisone and the 21-acylates thereof



United. States Patent 0' 1 r 2,841,600 6a-FLUORO A'-CORTISONE AND 6a-FLUORO A HYDROCORTISONE AND THE ZI-ACYLATES THEREOF John A. Hogg and George B. Spero, Kalamazoo Township, Kalamazoo County, Mich., assignors to The Upjohn Company, Kalamazoo, Mich., a corporation of No Drawing. Application January 18, 1957 0 Serial No. 634,828

4 Claims. (Cl. 260-397.45)

l LiAlHz 2,841,600 Patented July 1, 1958 This application is a continuation-in-part of application Serial No. 519,632, filed July 1, 1955.

It has been discovered that 6-fluoro-l-dehydrohydrocortisone and the 2l-esters thereof possess valuable antirheumatoid arthritic, anti-inflammatory and glucocorticoid activity. The compounds are useful in the treatment of inflammatory conditions of the skin, eyes and ears of humans and valuable domestic animals, contact dermatitis and other allergenic reactions. The compounds can be administered in conventional dosage forms such as pills, tablets and capsules for oral use or in conventional liquid forms as are used with natural and synthetic cortical steroid hormones for injection use. For topical use they can be administered in the form of ointments, creams, lotions and the like with or without coacting antibiotics, germicides and the like.

The compounds of this invention can be prepared in accordance with the following scheme of reactions:

H I H CH3 0H8 RI! I dH-o H (cHm-o CH3 CH3 ooon' 00012! on 111 II OHa OH:

III Ha CH8 CH3 cmon mime (:H OH H0 HO CH2 on W ,J V i 211:) ..-o i OH OH cnnro wherein R is fl-hydroxy or keto, R and R" are hydrogen or lower-alkyl, n is a whole number from one to two, X is fluorine, Ac is the acyl radical of a hydrocarbon carboxylic acid containing from one to twelve carbon atoms, inclusive, and R is hydrogen or Ac as defined above. The term lcwcr-alxyl, when used herein, refers to an all-:yl radical of from one to eight carbon atoms,

inclusive, e. g., methyl, ethyl, propyl, butyl, amyi, hexyl,

heptyl, ethylhexyl, and octyl.

Oxidation of the product, 6-fluoro-l-dehydrohydrocortisone 2l-acetate or other 2l-esters, e. g., with chromic acid in acetic acid, is productive of the Zl-esters of 6-fiuorod-dehydrocortisone, which compounds also possess glucocorticoid, antidnflarnmatory and anti-rheumatoid arthritic activity.

The above reactions as described hereinbelow are em bodiments of steps which will be exemplified in detail in the ensuing specification. it will be understood by those skilled in the art, nevertheless, that the specific order of steps may be inverted or transposed or otherwise varied to suit the purposes of economics, convenience, or the like.

The preferred compounds containing the 17 (20) double bond have the cis configuration, because the eis isomers can ultimately be converted in higher yields in the oxida tive 'hydroxylation step than is ordinarily possible with the trans isomer. it ShOU id be understood, however, that the trans isomer or mixtures of the cis and trans isomers can be used with satisfactory results.

In carrying out the epoxidation step shown above, a 3-ketal of 3-keto-11-oxygenated-4,17( 2())-pregnadiene-2=1- carbonyloxy steroid represented by Formula I, which can be prepared as disclosed in U. 5. Patent 2,707,184,

' preferably the 3-ethylene glycol ketal of methyl 3,11-diketo-4,17-(20) [cis] -'pregnadien-2 1-oate, is epoxidized With a peracid, e. g., peracetic or perbenzoic, or other known epoxidizing agents, to produce the corresponding 5:6-oxide (II). A mixture of both the :xand B-oxides is produced in this epoxidation reaction, and the mixture can be separated by chromatographic or crystallization techniques known in the art.

In the oxide opening step, a 3-ketalized 3-keto-5a,6..-.- oxido-l1-oxygenated 17(20) pregnene-Zl-carbonyloxy steroid (11), is reacted with hydrogen fluoride, to open the oxide ring and produce the corresponding 3-ketalized 3-keto-5hydroxy-6-fluoro-1l-oxygenated l7(20) -pregnene-Zl-carbonyloxy steroid (III). This epoxide opening step is ordinarily carried out at temperatures between about minus forty and plus fifty degrees ccntigradc, the preferred limits being between about zero and 25 degrees Centigrade. It can be performed under anhydrous condi tions in the presence or absence of a catalyst, e. g., boron 'trifiuoride; or under aqueous conditions in the presence or absence of a catalyst such as a hypohalous acid. Reaction conditions, for example, those disclosed by bchmid lin et 211., Helv. Chim. Acta, 36, 1241 ($53); Gallagher, J. Biol. Chem., 162, 495 (i946); Cornforth et at, J. Chem. Soc., 1954, 907 and Fried et al., J. Am. Chem. Soc, 75, 2273 (1953), are usually employed.

If anhydrous conditions are difficult or inconvenient to maintain, the oxide opening reaction can be performed under aqueous conditions in which case the ketal will be hydrolyzed at the same time. The thus-produced 3-!aeto group of Ila can then be reketalized in the manner described. hereinabove to produce the. corresponding ketal (III).

In the reduction step of the present invention, a 3- ketalized 3-keto 5 hydroxy-6-fluoro-l1-oxygenated-l7- (20)-pregnene-21-carbonyloxy steroid (III), preferably the 3-ethylene'glycol ketal of lower-alkyl, preferably methyl 3,1 1-diketo-5-hydroxy-6-1luoro-17 (20) -[cis]-pregnen-ZI-oate, is reduced with lithium aluminum hydride or other chemical carboxyl reducing agent in an organicv solvent, 'e. g., ether, dioxane, tetrahydrcfuran, benzene, to produce the corresponding 3-ketalized 5,11 3,2l-trihydroxy-6-fluoro-17(20)-pregnen-3-one. At completion of this reaction, the reactionrnixture is preferably mixed with water or, an acid, an ester or carbonyl agent followed by water, to decompose any excess lithium alumi num hydride and organo-"netal complexes. The usual reaction conditions for a lithium aluminum hydride reduction are employed, except that a reaction tempera: ture at room temperature or below is preferred, to ensure that reaction with the 6-fluoro does not occur. and acid, though operative and satisfactory under carefully controlled conditions, is preferably not employed in the decomposition step, to avoid undue hydrolysis of the ketal group.

The esterification step, i. e., to produce compound V, involves the conversion of a 2l-hydroxy group of 5,116,21-trihydroxy 6 fluoro-1700)-pregnen-3-one 3- alkylene kctal (IV) to a Zl-acyloxy group. Thisreaction can be performed under the esterification conditions known in the art, e. g., by the reaction of IV with the selected acid halide or acid chloride or acid bromide or the anhydride of a hydrocarbon carboxylic acid, or by reaction with the selected acid, in the presence of an esterification catalyst or with an ester under ester exchange reaction conditions. Reaction conditions which are apt to affect the labile lip-hydroxy group or 6-fiuoro group should be avoided. Compounds thus-produced include the compounds represented by Formula V wherein the 17(20)-configuration is cis, the 6-halogen is fluorine and the Ac is the acyl radical of a hydrocarbon carboxylic acid containing from one to twelve carbon atoms, inclusive, e. g., formic, propionic, butyric, isobutyric, valeric, isovaleric, trimethylacetic, Z-methylbutyric, 3-ethylbutyric, hexanoic, diethylacetic, triethylacetic, heptanoic, octanoic, a-ethylisovaleric, a cyclic acid, e. g., cyclopropylideneacetic, cyclopentylformic, cyclopentylacetic, fi-cyclohexylpropionic, cyclohexylformic, cyclohexylacetic, an aryl or alkaryl acid, e. g., benzoic, 2-, 3- or 4-rnethylbenzoic, 2,3-, 2,4-, 2,5-, 2,6-, '3,4- and 3,5-dimethylbenzoic, ethylbenzoic, 2,4,6-trimethylbenzoic, 2,4,6-triethylbenzoic, u-naphthoic, 3-methyl-a-naphthoic, an analkyl acid, e. g., phenylacetic, phenylpropionic, diphenylacetic, triphenylacetic, etc.

The oxidative hydroxylation of V to VI is carried out by reaction with a catalytic amount of osmium tetroxide and an oxidizing agent such as hydrogen peroxide, peracids, alkyl peroxides, amine oxide peroxides, and the like. A preferred procedure involves the use of'two to three molar equivalents of one of the oxidizing agents and less than 0.05 molar equivalent of osmium tetroxide, calculated on the basis of V, using tertiary butyl alcohol as the reaction medium and carrying out the reaction at about room temperature.

The hydrolysis step (VI- VII) involves the hydrolysis of the ketal group of the 3-ketalized 5,l1[3,17a-trihydroxy- 6-fluoro-21-acyloxypregnane-3,20-dione (VI), preferably the ethylene glycol ketal thereof, employing aqueous acid to produce the corresponding diketo compound VII. The hydrolysis is conveniently performed under relatively mild conditions, e. g., at room temperature with acetic acid or weak sulfuric acid.

The dehydration reaction is carried out using hydrogen chloride or Girards reagent T." Acetic anhydride, 'ptoluene-sulfonic acid and acetic acid, or thionyl chloride and pyridine are also suitable as dehydrating agents.

5 Care -shouldbe taken in this tion conditions that would affect the llfl-hydroxy group.

The l-dehydrogenatioii of VIII to IX (where R is a B-hydroxy group) can be carried out microbiologically,

i. e., fermentative l-dehydrogenation using Septomyxa or Fusarium organisms. If loss of the 2l-ester takes place during the fermentation, reacylation can be done by known '2l-acylation methods. Alternatively, the l-dehydrogenation can be carried out chemically in known manner by using selenium dioxide. 1

Alternatively, compound VIlIin the form of the 21- ester can'be oxidized to the corresponding ll-keto compound, i. e., l7a-hydroxy-6-fluoro-2l-acyloxy-3,11,20- trione (compound X), in accordance with the process for the oxidation of the 21-esters .of Kendalls compound F as disclosed in U. 5. Patent No. 2,751,402. Compound X can be subjected to l-dehydrogenation in the same manner as compound VIII thus producing l7ot-hydroxy-6- fiuoro-2l-acyloxy 1,4 pregnadiene-3,l1,20-trione (lX) Where R is a keto group.

Either of compounds VIII, IX or X can be transformed to the free 21-alcohol by hydrolysis in accordance with known methods for hydrolyzing compound F 21-esters to the free compound F alcohol. A preferred procedure 7 is to employ at least a'molar equivalent of an alkalimetal bicarbonate in a substantially oxygen-free solution of a mixture of a lower alkanol and water. The hydrolysis reaction is carried outat a temperature between ten and thirty degrees centigrade while protecting the mixture from atmospheric oxygen. After the hydrolysis is complete, the reaction mixture is neutralized with an acid, e. g., acetic acid, and the hydrolyzed product recovered from the reaction mixture. by evaporation and crystallization, extraction with methylene chloride, or the like. 7 7

As previously mentioned, the order of the various steps outlined above can be varied considerably. Suitable variations will occur to those skilled in the art, and the necessary adjustments can be readily appreciated and carried out by skilled chemists. For example, compound IV can be hydrolyzed to remove the 3-ketal, and this step followed in turn by 21-acylation, S-dehydration, oxidative hydroxylation and l-dehydrogenation under the above-described conditions.

The compounds represented by Formula IX and those represented by Formulas VIII and X, in addition to possessing the valuable anti-rheumatoid arthritic, antiinflammatory and glucocorticoid activity heretofore mentioned, are, additionally useful as intermediates for the preparation ofthe 9a-halo analogues thereof, namely v6 fiuoro 9a halo 1704,21 dihydroxy 4 pregnene- 3,11,20 trione, 6 fluoro 9a halo 11fl,l7a,21 trihydroxy 4 pregnene 3,20 dione including the cor responding 21- esters thereof; and 6-'fluoro-9ot-halo-17a,21- dihydroxy-1,4-pregnadiene-3,11,20-trione, v and 6-fluoroc halo 1lfi,17oc,2l trihydroxy 1,4 pregnadiene- 3,20-dione including thecorresponding 21-ester thereof. These compounds also possess the valuable anti-rheumatoid arthritic, anti-inflammatory and glucocorticoid activity of the parent 6-fluoro compounds, usually in higher degree. They possess a combination of high anti-inflammatory and glucocorticoid properties and at the same time possess low mineralocorticoid properties.

'In the preparation of these compounds, for example,

6- fluoro 11,8,17a dihydroxy 21 acetoxy 1,4 pregnadiene-3,20 dione of the class of Formula IX can be dehydrated withN-bromoacetamide and anhydrous sulfur dioxide until a negative acidified potassium iodide-starch test of the reaction mixture is obtained. Dilution with cold water, e. g., ice water, results in the precipitation of 6 fluoro 17a hydroxy 21 acetoxy-1,4,9(11)-pregnatriene-3,20-dione which can be purified with recrystallization from acetone. The crystalline product canthen be reacted in methylene chloride-tertiary butyl alcohol solution with perchloric acid and N-bromoacetamide or step to avoid various reac- N-iodosuccinimide to produce a reaction mixture from which 6-fluoro-9a-bromo-115, 17ot-dihydroxy-21-acetoxy- 1,4-pregnadiene-3,ZO-dione or the corresponding G-fluoro- 9et-iodo compound, respectively, can be recovered by precipitation with ice water and recrystallization from acetone. The latter compounds can be reacted in acetone solution with anhydrous potassium acetate at reflux temperature to produce 6-fluoro-9fi,1lfl-oxido-l7u-hydroxy 2l-acetoxy-l,4-pregnadiene-3,20-dione which is recoverable from the reaction mixture by chromatographic methods and can be purified by recrystallization from a Skellysolve B hexane hydrocarbons-acetone mixture. Reaction of the latter in methylene chloride solution with aqueous hydrogen fluoride at room temperature is productive of 9ot,6-difluoro-11p,17a-dihydroxy-2l-acetoxy- 1,4-pregnadiene-3,ZO-dione. Substitution of aqueous hydro gen chloride, and carrying out the reaction at lower temperatures, e. g., minus five degrees centigrade, is productive of 9ot-chloro-6-fluoro-11 3,17u-dihydroxy-21-acetoxy-l,4-pregnadiene-3,ZO-dione.

Substitution of 6-fluoro-11,8,17a-dihdroxy-2l-acetoxy- 4-pregnene-3,20-dione in the foregoing process is productive of the analogues resulting intermediate products, i. e., possessing the 3-keto-A -structure in the A-ring of the steroid molecule instead of the 3-keto-A -structure, and results in 9a,6-difiuoro-l15,17ot-dihydroxy2l-acetoxy-4-pregnene-3,ZO-dione and the corresponding 9achloro compound, respectively.

The compounds, 6-fiuoro-9ot-halo-17ot-hyd1'oxy-2l-acyloxy-4-pregnene-3,l1,20-trione (made in accordance with the process of the foregoing paragraphs, using as starting material 6-fiu0ro-17a-hydroxy-21-acyloxy-4-pregnene- 3,11,20-trione) and 6-fluoro-9a-halo-115,l7a-dihydroxy- 21-acyloxy-4-pregnene-3,ZO-dione, can be utilized as starting material for the corresponding 6-fluoro-9a-l1alo- 17a hydroxy 21 acyloxy 1,4 pregnadiene 3,11,20- trione and 6-fiuoro-9a-halo-11B,17a-dihydroxy-21-acyloxy-l,4-pregnadiene-3,20-dione by fermentative l-dehydrogenation using Septomyxa or Fusarium organisms or by chemical l-dehydrogenation using selenium dioxide under the conditions previously described for the l-dehydrogenation of compound VIII to compound IX.

The 90,6 difluoro 1113,1711 dihydroxy 21 acetoxy- 1,4pregnadiene-3,ZO-dione or the 9a-chloro analogue thereof can be oxidized with chromic acid in accordance with known methods for converting Kendalls compound F 2l-esters to Kendalls compound E 21-esters to produce 90,6 difluoro 17a hydroxy 2l-acetoxy-l,4-pregnadiene-3,11-20-trione and 9a-chloro-6-fiuoro-17u-hydroxy-Z1-acetoxy-1,4-pregnadiene-3 ,l1,2 -trione, tively. In a similar manner, the above compounds possessing the 3-keto-A -structure in the A-ring can be oxidized with chromic acid to produce 9a,6-diiluoro-17mhydroxy 21- acetoxy 4 pregnene 3,11,20 trione and 9a chloro 6 fluoro 17oz hydroxy 21 acetoxy- 4-pregnene-3,1 1,20-trione, respectively.

The foregoing 6-fiuoro-9a-halo compounds can be hydrolyzed to the corresponding 21-hydroxy compounds according to known methods for hydrolyzing compound F 21-acetate to compound F, for example, in aqueous solution with potassium bicarbonate under oxygen-free.

form. The foregoing process produces a mixture in which.

the 6 3-form predominates. The 6a-epimer can be separated from the products by chromatographic or fractional crystallization techniques known in the art. The

respeccompounds are useful, however, as such in the mixtures as produced directly from the synthesis steps described. It is occasionally desirable, nevertheless, to produce from the mixture by separation or by epimerization the 6aepimer which possesses even higher activity than the 6pepimer. Conversion of the 6,6-epimer or mixtures predominating therein can be accomplished by treatment at temperatures of zero degrees centigrade or slightly below in an essentially anhydrous liquid medium with an an-' hydrous acid, such as hydrochloric acid. The mixture should be maintained at temperatures below zero degrees Centigrade, or at least below room temperature during the addition of the acid. The reaction mixture can then be washed with successive portions of dilute alkali and water, and then dried and evaporated under reduced pressure. The 6a-fiuoro products can be recovered from the crude reaction product and purified by recrystallization.

The following examples are illustrative of the process and products of the present invention, but are not to be construed as limiting.

EXAMPLE 1 The 3-etlzylent glycol ketal of methyl 3,1I-d"'keto-5a,6a oxide-1 7(20 [cis] -pregnen-21-oate To a solution of 5.0 grams of the I i-ethylene glycol ketal of methyl 3,1l-diketo-4,17(20)-'[cis]-pregnadien 21-oate, prepared in the manner described in U. S. Patent 2,707,184, in milliliters of chloroform Was added a chilled solution of 1.9 grams of perbenzoic acid dissolved in 31.5 milliliters of chloroform. The solution was maintained at about .four degrees centigrade for 24 hours, and then at room temperature for 72 hours. The solution was then washed with a live percent aqueous solution of sodium bicarbonate and then with water. The chloroform layer was separated, dried and the solvent distilled to give a residue of 5.3 grams of solid. Crystallization of this solid from methanol gave 2.24 grams of product melting at 'to degrees centigrade and after two crystallization's from ethanol, there was obtained pure 3-ethylene glycol ketal of methyl 3,11-diketo-5a,6ot-oxido- 17(20)-[cis]-pregnen21-oate melting at 206 to 209 degrees Centigrade having an [cab of 37 degrees (CHCI and having the analysis given below:

Calculated for C H O C, 69.20; H, 7.75. C, 69.59; H, 7.81.

Found:

EXAMPLE 2 Methyl 3,1 Z-diket0-5ot,6{3-dihydr0xy-l 7(20) -all0pragnen- 21-0ate and methyl 3,] 1-diket0-5u-hydr0xy-6p-flu0r0 I 7(20 -all0pregizen-21-0ute To a solution of 1.73 grams of 3ethylenc glycol ketal of methyl 3,11-dil eto-5a,6u-oxido-17(20)-[cisl-pregnen- 2l-oate in sixteen milliliters of methylene chloride was added six milliliters of 48 percent hydrofluoric acid. The heterogeneous mixture was stirred for two hours, made slightly basic with 300 milliliters of five percent sodium bicarbonate solution, and extracted with methylene chloride. The extract was washed, dried, and evaporated to dryness to give 1.62 grams of crude solid. Chromatography gave two fractions: A, 481 milligrams eluted with methylene chloride plus five percent acetone and B, 921 milligrams eluted with methylene chloride plus ten and twenty percent acetone. Crystallization of fraction A from acetone-Skellysolve B hexanes gave 390 milligrams of methyl 3,1 1-dilteto-5whydroxy-6,8-fluoro-17(20) allopregnen-Zl-oate, melting point 254 to 260 degrees centigrade. The analytical sample melted at 260 to 263 degrees centigrade.

F Anajlysis-Calculated for C H O F: F, 4.84. Found:

Fraction B on crystallization from"acetone-Skellysolve B hexanes gave 470 milligrams of methyl 3,11-diketo- 5ct,6/3 dihydroxy 17(20) allopregnen 21 oate.

' or the like.

, EXAMPLE 3 Methyl 3,11-diketo-5a-hydr0xy-6B-fluor0-17(20)-all0- pregnen-ZJ-oate 3-ethylene ketal A mixture of 1.9 grams of methyl 3,11 diketo-5ahydroxy-6fi-fluoro-17(20)-allopregnen-21oate, 59 milligrams of p-toluene-sulfonic acid monohydrate and 31 milliliters of distilled ethylene glycol was added to 800 milliliters of benzene. The mixture was stirred and refluxed for two hours, with the condensate passing through a water trap to remove the water. After reflux the mixture was cooled, washed with water and evaporated to dryness to give a crude solid which on recrystallization from acetone-Skellysolve B hexanes gave 1.96 grams of methyl 3,11-diketo-5a-hydroxy-6fl-fiuoro-17(20)-allopreg-- nen- Zl-oate 3 ethylene ketal, melting point 170 to 173 degrees centigrade. v

Following the above procedure, substituting other dihydric alcohols for ethylene glycol, for example, 1,2- propylene glycol, 2,3-butane-diol, 1,3-butanediol and 2,3- pentanediol is productive of the respective 3-alkylene ketals of methyl 3,11 diketo 5a hydroxy 6,8 fluoro- 17 (20) allopregnen-Z l-oate.

EXAMPLE 4 5 ,1 1 3,21 -trihydrxy-6B-flu0r0-1 7 (20) -al[opregnen-3-one 3-ethylene ketal To a solution of 1.96 grams of methyl 3,11-dike-to-w 'hydroxy-6B-fluoro-17 (20) -allopregnen-21-oate 3-ethylene ketal in 850 milliliters of anhydrous ether was added 3.7 grams of lithium aluminum hydride and the mixture was stirred fora period of one hour. 200 milliliters of water was added slowly and the ether phase separated. The

The acetate was prepared by allowing 0.87 gram of 5u,l1/3,21 trihydroxy 6,3 fluoro 17 (20) allopregnen-3-one 3-ethylene ketal to standovernight in ten milliliters of aceticanhydride and ten milliliters of pyridine. The solution was then poured into ice water to give 0.92 gram of 5u,1lB-dihydroxy-6B-fiuoro-21-acetoxy-17(20)- allopregnen-3-one 3-ethylene ketal, melting point 140 to 150 degrees centigrade, which on recrystallization from acetone-Skellysolve B hexanes gave 0.77 gram, melting point 149 to 153 degrees Centigrade.

Similarly, other Zl-organic carboxylic esters of Scull 3,21 trihydroxy 6p fluoro 17(20) allopregnen- 3-one 3-ethylene ketals are prepared wherein the 21- acyloxy group is formyloxy, propionyloxy, butyryloxy, valeryloxy, hexanoyloxy, heptanoy-loxy, octanoyloxy, benzoyloxy, phenylacetoxy, or the like, by contacting 504,113,2 1 trihydroxy 6,8 flucro 17(20) pregnen 3- one 3-ethylene ketal with an appropriate acylating agent, e. g., the anhydride or acid halide of the selected acid in a solvent such as, for example, benzene, toluene, acetic acid,

A convenient method of preparing the 21- forrnyloxy ester consists in contacting 5a,11,8,21-trihyt0 droxy-6,8-fiuoro-17(20)pregnen-3-one 3-ethylene ketal with formic acid in the presence of para-toluenesulfonic acid. j EXAMPLE 6 5 00,1 15,1 7wtrihydr0xy-6B-flu0r0-21 acetoxyallop regitane- 3,20-di0ne 3-ethylene ketal i To a solution of 0.77 gram of 5a,11;8-dihydroxy-6B- fiuoro-21-acetoxy-l7(20)-allopregnen-3 one 3-ethylene ketal in 35 milliliters of tertiary butyl alcohol was added one milliliter of pyridine, 1.9milliliters of N-methylmorpholine oxide peroxide solution, and 13.1 milligrams of osmium tetroxide (9.1 milliliters of tertiary butyl alcohol solution containing 1.44 milligrams osmium tetroxide per milliliter). The solution was stirred for a period of 2.5 hours, fifteen milliliters of five percent sodium hydrosulfite added, stirred for an additionalten minutes, 0.7 gram of finely ground synthetic magnesium silicate added, stirred for a period of twenty minutes more and filtered. The filtrate was taken to dryness under reduced pressure (below fifty degrees centigrade) and the residue dissolved inmethylene chloride, washed with water, dried and evaporated to dryness. This residuewas crystallized from acetone-Skellysolve B hexanes to give 0.47 gram of 504,11B,17a-trihydroxy-6B-fiuoro-2l acetoxyallopregnane- 3,20-dione 3-ethylene ketal, melting point 220 to 228 degrees centigrade.

, EXAMPLE 7 5 0a,] 15,17a-trihydroxy-tiB-fluoro-ZI acetoxyallopregnane- 3,20-di0ne V A solution of 0.47 gram of 5u,1118,17a-trihydroxy-6flfluoro-Z1-acetoxyallopregnane-3,20-di0ne B-ethylene ketal in 35 milliliters of acetone and four milliliters'of 1 N sulfuric acid solution was gently boiled on the steam bath for ten minutes, cooled and neutralized with dilute sodiumrbiearbonate solution. Addition of water and cooling gave 0.33 gram of 5a,11B,17a-trihydroxy-6fifluoro-Zl-acetoxyallopregnane-3,20-dione, melting point 230 to 240 degrees Centigrade.

EXAMPLE 8 6B-fluor0-1 113,1 7a-dihydroxy-21 -acet0vcy-4-pregnene-3,20- dione (6p-fluorohydroco rtisone acetate) EXAMPLE 9 A medium consisting of one percent dextrose hydrate, two percent cornsteep liquor of sixty percent solids and Kalamazoo tap water was adjusted to pH 4.9 sodium hydroxide. The medium was steam sterilized at fifteen pounds pressure for thirty minutes, cooled, and then inoculated with a 24-hour growth, from spores, of Septomyxa afiinis, A. T. C. C. 6737. The medium was agitated, sparged with sterile air at the rate of one-tenth volume of air per volume of medium per minute. At the end of 24 hours of fermentation at room temperature, the pH was about 7.4. To this culture there was added a solution of 6,8-fluoro-11p,17a-dihydroxy-21-acetoxy-4- pregnene-3,20-dione (6,8-fiuorohydrocortisone acetate),

Infrared data and ultraviolet data are in dissolved in diethylforrnamid The solution was prepared by dissolving five parts of the steroid in 100 parts of the solid and adding about ten centimeters of the solution per liter of the medium. Fermentation was continued for a period of 48 hours whereupon the mycelium and beer were extracted thoroughly with methylene chloride. The extract was washed with sodium bicarbonate solution and then with water, dried and concentrated in vacuo to a slightly viscous residue. The residue, alter reacetylation with acetic anhydride in pyridine, was fractionated chroniatographically and 65-iluoro-115,l7adihydroxy-21-acetoxy-1,4-pregnadiene-3,ZO-dione was recovered as a light-colored crystalline solid.

Following the procedure of Example 9, above, but substituting as starting material 9a,6-difiuoro-l1,6,17e-dihydroxy-21-acetoxy-4-pregnene-3,ZO-dicne and reacetylating the recovered crude product, there was produced 904, 6-difiuoro-1lfi,17u-dihydroxy-21-acetoxy-1,4-pregnadiene- 3,20-dione as a white crystalline solid.

EXAMPLE Following the above procedure, but replacing the 6,8- fiuoro-l1,6,17a-dihydroxy-21-acetoxy-4-pregnene 3,20-dione (6B-fluorohydrocortisone acetate) by 6,8-fluoro-17ahydroxy-21-acetoxy-4-pregnene-3,11,20-trione (6fi-fluorocortisone acetate) which was made by the oxidation of the product of Example 8 according to the methods disclosed in U. S. Patent 2,751,402, yielded 6(3-fluoro-17a hydroxy-2l-acetox-1,4-pregnadiene-3,lLZS-trionc as a crystalline product.

As previously described, the 21-acyloxy products of this invention, e. g., the products of Examples 9 and 10 can be hydrolyzed to the corresponding free 21-alcohols by the hydrolysis procedure described above. In the hydrolysis procedure, as previously stated, it is desirous to employ an oxygen-free solution and oxygen-free conditions and to use a molar excess of an alkali-metal bicarbonate such as potassium bicarbonate as a hydrolytic agent. The temperature is preferably held lower, i. e, between ten and about thirty degrees Centigrade, and the hydrolyzed solution subsequently neutralized with an acid such as acetic acid.

The compounds disclosed in Examples 9 and 10 possess valuable anti-rheumatoid arthritic, anti-inflammatory activity and can be used and administered as disclosed in the foregoing description.

EXAMPLE ll lsomerization of 6,6- to 6a-flu0rchyclrocortisone 21 -acetare A solution of 0.132 ram of 6fi-fluorohydrocortisone 21-acetate in twelve milliliters of chloroform and 0.1 milliliter of absolute alcohol was cooled to minus ten degrees centigrade in an ice-salt bath and a stream of anhydrous hydrochloric acid was gently bubbled through the solution for 2.5 hours while the temperature was maintained between minus five and minus fifteen degrees centigrade. The solution was then diluted with 25 milliliters of chloroform, washed with dilute sodium bicarbonate and water, dried over anhydrous sodium sulfate, and

evaporated to dryness under reduced pressure at sixty degrees centigrade or less. Crystallization of the residue from acetone-Skellysolve B gave 42 milligrams of product, 6u-fluorohydrocortisone ZI-acetate, melting point 203 to 210 degrees centigrade.

in a similar manner, 6fi-fiuoro-11/3,17a-dihydroxy-21- acetoxy-1,4-pregnadiene-3,20-dione is converted to fiuoro-l1B,17tt-dihydroxy 21 acetoxy-l,4-pregnadiene- 3,20-dione, melting point 238 to 242 degrees Centigrade,

pregnadiene-3,20-dione is converted to 6u,9a-difluoro- 116,17a-dihydroxy 21 acetoxy-l,4-pregnadiene-3,20 dione.

It is to be understood that the invention is not to be iimited to the exact details of operation or exact compounds shown and described, as obvious modifications and equivalents will be apparent to one skilled in the art, and the invention is therefore to be limited only by the scope of the appended claims.

We claim:

1. 6oc-flu0r0 11 oxygenated 17ot,21 dihydroxy-1,4- pregnadiene-3,20-dione of the following formula:

wherein R is a member of the group consisting of 5- hydroxy and keto; and the 21-acylates thereof wherein the acyl radical is that of a hydrocarbon carboxylic acid containing from one to twelve carbon atoms, inclusive.

2. 6a-fiuoro-l-dehydrohydrocortisone.

3. 6a-fluoro-l-dehydrocortisone.

References Cited in the file of this patent UNITED STATES PATENTS 2,705,237 Djerassi Mar. 29, 1955 2,756,179 Fried July 24, 1956 2,769,823 Schneider Nov. 6, 1956 OTHER REFERENCES Mattox: Journal of Biological Chemistry 197 (1952), pp. 261-270.

Herzog: I. A. C. S., 77, 4781 (1955).

Hogg: I. A. C. S., 77, 4438 (1955).

Burstein: J. A. C. 8., 77, 4668 (1955).

Szpilfogel: Rec. Trav. Chirn., 75, 475 (1956).

Herzog: I. A. C. 5., 79, 502-3 (1957).

UNITED STATES PATENT OFFICE Certificate of Correction Patent No. 2,841,600 July 1,1958 John A. Hogg et a1.

It is hereby certified that error appears in the printed specification of the above numbered patent requiring correction and that the said Letters Patent should read as corrected below.

Columns 1 and 2, Formula I should appear as shown below instead of as in the patent c o o R n 511-0 oncolumn 2, Formula He should appear as shown below instead of as in the patent:

columns 1 and 2, Formula III should appear as shown below instead of as in the patent:

column 2, Formula V should appear as shown below of as in the patent:

column 2, strike out the arrow from compound I to compound III; same column, insert an arrow from compound II to compound IIa; same column 2 insert LiAlI-L- between compound III and. compound IV; column 1, strike out LIriAlH Signed and sealed this 13th day of October 1959 Attest: KARL H. AXLINE, Attestz'ng Oyfieer.

ROBERT C. WATSON, Commissioner of Patents. 

1. 6A-FLUORO - 11 - OXYGENATED - 17A,21 - DIHYDROXY-1,4PREGNADIENE-3,20-DIONE OF THE FOLLOWING FORMULA: 